应用荧光定量聚合酶链方法检测宫颈组织中人乳头瘤病毒16E6基因  

Detection of HPV16 E6 gene in cervical tissues by quantitative polymerase chain reaction

作　　者：马彩玲[1] 李轶杰[2] 张富春[2] 王国荃[3] 郑玉建[4] 开丽曼[1] 热西旦[2] 韩英[1] 帕提古丽 

机构地区：[1]新疆医科大学第一附属医院妇产科,乌鲁木齐830054 [2]新疆大学生命科学与技术学院分子生物学重点实验室,新疆生物资源基因工程重点实验室 [3]新疆医科大学公共卫生学院环境卫生教研室 [4]不详 [5]新疆喀什地区疏勒县人民医院妇产科

出　　处：《中华医学杂志》2004年第6期469-473,共5页National Medical Journal of China

基　　金：国家自然科学基金资助项目 (3 9960 0 79)

摘　　要：目的　建立荧光定量聚合酶链 (FQ PCR)方法检测宫颈细胞中人乳头瘤病毒 16型(HPV16 )E6基因的方法。方法　以 2 2 0例新疆妇女宫颈不同病变组织DNA作为样本 ,人乳头瘤病毒 16型 (新疆株 )E6基因 (HPV16E6 ,AF32 785 1)作为扩增的靶基因 ,同时以人 β 肌动蛋白基因片段作为内参照 ,借助于设计的目的基因引物 ,两个特异的荧光探针 ,在筛选的FQ PCR优化反应体系中 ,同时对两个基因片段进行扩增 ,得到HPV16E6的相对含量。结果　对 β 肌动蛋白阳性的宫颈不同病变组织中HPV16E6阳性率及其相对含量进行统计分析 ,新疆妇女宫颈脱落细胞标本 96例中 ,HPV16E6阳性 3例 (3 3% ) ;宫颈癌蜡块组织 5 9例中HPV16E6阳性 4 9例 (83 0 % ) ;宫颈上皮内瘤变 (CIN)蜡块组织 6 5例中 ,HPV16E6阳性 2 8例 (75 7% ) ;宫颈炎蜡块组织 33例 ,HPV16E6阳性 2 8例 (93 3% )。HPV16E6的相对含量随病情加重呈上升趋势 ,二者呈显著正相关 ,相关系数为 0 83。结论　建立的FQ PCR检测宫颈不同病变组织内HPV16E6基因的方法 ,能反映单位细胞人乳头瘤病毒的复制情况 ,筛查宫颈癌患者 ,并可能应用于宫颈癌的风险评估和疗效观察。Objective To establish a method for detection of Human Papillomavirus (HPV) type16 E6 gene in Cervical carcinomas Specimens. To study the relationship between the quantities of HPV16 E6(Human papillomavirus type16 E6 gene) in cervical tissues and the course of cervical disease in Xinjiang. Methods HPV16E6 gene and β-actin was detected in parallel by FQ-PCR (fluorescence quantitative PCR). The number of copies of HPV16 E6 gene and β-actin was detected in parallel by FQ-PCR(fluorescence quantitative PCR ) in tissues of 69 cervical cancer, 65 cervical intraepithelial neoplasia (CIN), 33chronic cervicitis and samples of 96 cervical smear samples of vaginitis and cervicitis. The variation in HPV copies per genomic DNA equivalent can be estimated by dividing the HPV copy number by the β-actin copy number. Results The positive rate of HPV16 E6 gene was 83.0%,75.7%,93.3% and 3.3% in tissues of cervical cancer, cervical intraepithelial neoplasia (CIN), chronic cervicitis and samples of cervical smear respectively. The amount of HPV16 E6 gene was gradually higher by the developing of the course of cervical disease. They have positive rank correlation, r=0.83 , P <0.01. Conclusion The study underscores the importance of the relationship between the HPV16 E6 gene and the course of cervical disease in Xinjiang.It also suggests that the quantification of HPV16 E6 gene may be useful as a prognostic tool to identify women who are at increased risk of developing cervical cancer. This method may be applied to studies of a number of issues related to the natural history of cervical cancer, such as the amounts of HPV in high- and low-grade lesions.

关 键 词：荧光定量聚合酶链法 宫颈肿瘤 人乳头瘤病毒16 E6基因 脱落细胞学检查 

分 类 号：R737.33[医药卫生—肿瘤] R730.43[医药卫生—临床医学]